Biological agent detector method

ABSTRACT

A biological agent detector detects the presence of any biological agents, such as anthrax or other biological warfare agents, in a sample of air. The biological agent detector includes a bio-concentrator that concentrates an aerosol and a pyrolyzer portion including two detecting devices. One detecting device operates in a sample collection mode and collects a sample of air when the other detecting device operates in a sample analysis mode and analyzes a sample of air. After a predetermined amount of time, the detecting devices switch functions, providing continuous sampling of air.

REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 11/174,925 filed on Jul. 5, 2005, now U.S. Pat. No. 7,837,937 issued on Nov. 23, 2010, which claims priority to U.S. Provisional Application No. 60/586,902 filed on Jul. 9, 2004.

BACKGROUND OF THE INVENTION

The present invention relates generally to a biological agent detector including two detecting devices that each alternate between a sample collection mode and a sample analysis mode to provide continuous aerosol collection and air monitoring for biological agents.

As an example biological agent detector, a Chemical Biological Mass Spectrometer Block II (CBMS II) detects biological agents present in the air by employing a two-step process using a detector including a pyrolyzer and an associated mass spectrometer. During sample collection, a first pump draws 330 liters of air per minute into an inlet of a bio-concentrator. The bio-concentrator extracts aerosol particles that are drawn into the pyrolyzer by a second pump at 1 liter per minute. The aerosol drawn into the pyrolyzer includes secondary particles, such as dust, fibers, and dirt, and may also include biological agents, such as anthrax or other biowarfare agents. A sample of the aerosol collects at the bottom of a pyrotube in the pyrolyzer. The air and aerosol particles not collected in the pyrotube are exhausted through an exhaust outlet.

During sample analysis, the aerosol sample collected at the bottom of the pyrotube is analyzed to identify any biological agents in the air. A small droplet of a methylating reagent, such as tetramethylammonium hydroxide (TMAH) dissolved in methanol, is added to the air sample with a hypodermic needle. The hypodermic needle is typically made of stainless steel. If the sample includes any biological agents, the methylating reagent derivatizes organic materials to make them more volatile. For example, fatty acids in the cell walls of the bacterial agents form Fatty Acid Methylated Esters (FAMEs).

The sample in the pyrotube is then rapidly heated or pyrolyzed to form gas molecules during an analysis phase. A third pump draws 1 milliliter per second into the mass spectrometer for identification of any biological agents. Biological agents have distinctive patterns of organic molecules. If any biological agents are present in the sample, the mass spectrometer identifies the biological agents by determining the molecular pattern. After a molecular pattern is determined, the pattern is compared to a pre-programmed list of patterns for identification of any biological agents in the air sample.

A drawback to the prior art biological agent detector is that the prior art system must stop collection of an aerosol sample in the pyrotube during the analysis phase, and therefore does not detect any biological agents present in the air during sample analysis. Therefore, the sample collection is intermittent, with time gaps in which no air monitoring occurs. This increases the time required to detect the presence of any biological agents and in some situations may result in a failure to detect a transient agent.

To provide continuous detection and analysis, a second pyrolyzer and bio-concentrator are needed. However, duplicating the bio-concentrator and the pyrolyzer is expensive.

Thus, it is desirable to have a biological agent detector including two detecting devices that share a bio-concentrator and that each alternate between a sample collection mode and a sample analysis mode to provide continuous collection and monitoring of an air sample, as well as overcoming the other above-mentioned deficiencies of the prior art.

SUMMARY OF THE INVENTION

A biological agent detector detects the presence of any biological agents, such as anthrax or other biological warfare agents, in a sample of air. The biological agent detector includes a bio-concentrator that concentrates an aerosol within the sample of air and a pyrolyzer portion including two detecting devices. One detecting device operates in an aerosol sample collection mode and extracts the aerosolized particles from the sample of air when the other detecting device operates in a sample analysis mode and analyzes the particles collected from a previous sample of aerosol. After a predetermined amount of time, the detecting devices switch functions.

The bio-concentrator draws air into the biological agent detector through an inlet and separates aerosol particles from the main airflow. The bulk of the air is exhausted through an exhaust. The bio-concentrator is moveable so that a concentrated aerosol outlet of the bio-concentrator is alignable with an inlet of each of the detecting devices. Physically moving the bio-concentrator so that the concentrator aerosol outlet aligns with the inlets of each of the detecting devices permits an uninterrupted flow of concentrated aerosol from the bio-concentrator to the detecting device with no sharp bends or discontinuities in the flow path.

When the concentrated aerosol outlet of the bio-concentrator is aligned with a first detecting device, the sample concentrated in the bio-concentrator flows along a stainless steel inlet tube and collects in a closed end quartz pyrotube in the first detecting device.

When the air sample is being collected in the first detecting device, an air sample previously collected in the second detecting device is analyzed. A small droplet of a methylating reagent, such as 0.0015 Molar to 0.015 Molar TMAH dissolved in methanol, is added to the sample collected in the pyrotube. The methylating reagent splits open the cells of any bacterial agents in the sample and neutralizes and derivatizes the organic molecules in the agents to more volatile compounds. The sample is also heated to form pyrolysis products.

The pyrolysis products are transported from the pyrotube to a mass spectrometer for identification and analysis. The mass spectrometer includes a preprogrammed list of molecular patterns that are each associated with a specific biological agent. If any biological agents are present the sample, the mass spectrometer identifies the biological agents by comparing the identified molecular pattern to the list of molecular patterns preprogrammed into the mass spectrometer. After a match is found, the mass spectrometer identifies the associated biological agent.

After a predetermined amount of time, the functions of the detecting devices are switched. The bio-concentrator moves so that the concentrated aerosol outlet is aligned with the inlet of the second detecting device for collection of an aerosol sample. Therefore, the biological agent detector continuously collects an air sample for analysis.

These and other features of the present invention will be best understood from the following specification and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The various features and advantages of the invention will become apparent to those skilled in the art from the following detailed description of the currently preferred embodiment. The drawings that accompany the detailed description can be briefly described as follows:

FIG. 1 illustrates a bio-concentrator and an aerosol flow switch mechanism of a biological agent detector;

FIG. 2 illustrates a front view of a pyrolyzer portion of the biological agent detector including two detectors;

FIG. 3 illustrates an enlarged front view of the pyrolyzer portion of FIG. 2; and

FIGS. 4A and 4B illustrates movement of bio-concentrator from a first position when it aligns with one detecting device to a second position where it aligns with the other detecting device.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

FIG. 1 illustrates a bio-concentrator 22 and an aerosol flow switch mechanism 23 of a biological agent detector 20 of the present invention. The biological agent detector 20 detects the presence of any biological agents, such as anthrax or other biological warfare agents, in a sample of air.

The bio-concentrator 22 concentrates an aerosol from an air stream and provides the aerosol to one detecting device 26 a and 26 b of a pyrolyzer portion 21 for collection and analysis. The aerosol flow switch mechanism 23 provides the aerosol to one of the detecting devices 26 a and 26 b, while the aerosol sample in the other of the detecting devices 26 a and 26 b is analyzed.

The bio-concentrator 22 draws over 300 liters of air per minute into the biological agent detector 20 through an inlet 71 (shown in FIGS. 4A and 4B). The bio-concentrator 22 separates aerosol particles from the main airflow and exhausts the bulk of the air through an exhaust 73 and an aligned exhaust port 95 at approximately 299 liters/minute. The bio-concentrator 22 provides a concentrated airflow of approximately 1 liter per minute containing almost the entire aerosol of interest through a concentrated aerosol outlet 62 that leads to one of the detecting devices 26 a and 26 b for collection. The concentrated airflow contains about 90% of the bio-particles.

The bio-concentrator 22 is clamped on a turntable 60 rotatable about a pivot 88. The bio-concentrator 22 is moveable between two positions. The concentrated aerosol outlet 62 is alignable with one detecting device 26 a in a first position and the other detecting device 26 b in a second position. The air stream from the concentrated aerosol outlet 62 flows through a small hole 110 in the turntable 60 that is aligned with the concentrated aerosol outlet 62. The turntable 60 rests on and is rotatable relative to a stainless steel base plate 66.

The base plate 66 includes two holes 112 a and 112 b that are each aligned with one of the detecting devices 26 a and 26 b, respectively. When the turntable 60 rotates, the small hole 110 in the turntable 60 aligns with one of the holes 112 a and 112 b in the base plate 66. The sidewalls of the holes 112 a and 112 b are polished to eliminate the possibility of discontinuities in the walls of the aerosol path. An upper surface 118 of the base plate 66 includes a groove that surrounds each of the holes 112 a and 112 b, and a rubber o-ring 114 a and 114 b is received in each of the grooves to provide a relatively air tight seal between the upper surface 118 of the base plate 66 and the lower surface 116 of the turntable 60.

The base plate 66 includes hardened steel pins that penetrate grooves in the turntable 60 to limit rotation of the turntable 60 and to provide a precise stop so that the hole 110 in the turntable 60 aligns with one of the two holes 112 a and 112 b in the base plate 66. A compression spring 72 (shown in FIGS. 4A and 4B) provides 10 pounds of force between the turntable 60 and the base plate 66 to ensure that the o-rings 114 a and 14 b are compressed and that the lower surface 116 of the turntable 60 contacts the upper surface 118 of the base plate 66. The precise alignment stops and the compression force ensures a smooth path for the aerosol stream with minimum discontinuities in the sidewalls of the holes 112 a and 112 b.

The upper surface of the turntable 60 includes a conical alignment feature 64 feature that mates with a conical aperture 130 in the lower surface of the bio-concentrator 22 to ensure that the concentrated aerosol outlet 62 of the bio-concentrator 22 is precisely aligned with the through hole 110 in the turntable 60. A locking mechanism 99 locks the bio-concentrator 22 in position.

As shown in FIGS. 4A and 4B, a motor 70 with a reduction gearbox, a crank and a pushrod rotates the turntable 60 approximately 180° between the first position and the second position. Micro-switches detect the position of the crank. The crank rotates to move the turntable 60 so that the concentrator exhaust outlet 62 of the bio-concentrator 22 precisely aligns with one of the detecting devices 26 a and 26 b. A solenoid lifts the turntable 60 from the base plate 66 and the o-rings 114 a and 114 b before the turntable 60 moves to eliminate wear on the o-rings 114 a and 114 b. When the hardened steel pin precision end stops prevent the turntable 60 from moving, the solenoid is deactivated and the compression spring 72 pushes the turntable 60 back down against the base plate 66. The mechanism that switches the bio-concentrator 22 outlet aerosol flow between the two detecting devices 26 a and 26 b operates without compromising the aerosol concentration. To efficiently transport the aerosol, several precautions must be taken. The airflow path must be as smooth as possible. Unnecessary turbulence will cause the particles to be thrown against the sidewalls of the transport tubing. Sharp turns in the particle flow path will also cause the particles to be impacted against the sidewalls.

FIG. 2 schematically illustrates the pyrolyzer portion 21 of the biological agent detector 20 and an interface 22 a with the bio-concentrator 22. The pyrolyzer portion 21 includes a body portion 24 and support rods (not shown) that support the body portion 24. The pyrolyzer portion 21 includes the two detecting devices 26 a and 26 b. In one example, the detecting devices 26 a and 26 b are pyrolyzers. The body portion 24 includes an embedded heater and temperature sensor device 28. The pyrolyzer portion 21 are attached to an underside of the base plate 66 such that the moveable bio-concentrator 22 is located above the base plate 66.

Each detecting device 26 a and 26 b includes a stainless steel inlet tube 30 a and 30 b, respectively. Each inlet tube 30 a and 30 b includes a gradually tapered adapter 68 a and 68 b that increases the airflow velocity in the inlet tubes 30 a and 30 b. The inlet tubes 30 a and 30 b are made of an electrically conducting material that is electrically grounded to prevent electrostatic buildup. The inlet tubes 30 a and 30 b are electro-polished to smooth and passivate the internal surfaces. The inlet tubes 30 a and 30 b are not an integral part of the body portion 24 and can be removed for cleaning and replacement.

The sample air exits the inlet tubes 30 a and 30 b through an outlet 32 a and 32 b, respectively, and collects in a respective closed end quartz pyrotube 34 a and 34 b. The pyrotubes 34 a and 34 b each have a substantially conical bottom that provides a small collection volume for the sample. The diameter of the inlet tubes 30 a and 30 b forces approximately 1 liter per minute of aerosol flow to form a jet that impacts the internal vertex of the pyrotube 34 a and 34 b. The particles carried by the airflow are impacted against the internal closed end of the pyrotube 34 a and 34 b whereas the air is exhausted. The efficiency of particle impaction and collection requires close tolerances between the tip of the inlet tubes 30 a and 30 b and the bottom of the pyrotubes 34 a and 34 b.

Each detecting device 26 a and 26 b includes a wide stainless steel aerosol outlet exhaust line 36 a and 36 b that carries an exhaust flow at approximately 1 liter per minute. The wide diameter of the exhaust line 36 a and 36 b prevents fibrous and other particles that fail to impact and collect at the closed end of the pyrotube 34 a and 34 b from building up and clogging the exhaust lines 36 a and 36 b.

A injector needle 46 a and 46 b adds a small droplet (1 to 2 microliters) of a methylating reagent from a reagent source 44 a and 44 b (shown in FIGS. 4A and 4B) to the sample collected in the pyrotube 34 a and 34 b. In one example, the methylating reagent is 0.0015 Molar to 0.015 Molar TMAH dissolved in methanol. The injector needles 46 a and 46 b are positioned to deliver a methylating reagent droplet to the bottom-center of the respective pyrotube 34 a and 34 b. The methylating reagent splits open the cells of any bacterial agents in the sample and neutralizes and derivatizes the organic molecules in the agents to more volatile compounds. The injector needles 46 a and 46 b are a fused quartz capillary tube. The quartz is inert and resistant to the thermal cycling of the detecting devices 26 a and 26 b and the corrosive methylating reagent.

Each detecting device 26 a and 26 b is fitted with an injector needle port that seals the injector needle 46 a and 46 b at the point of entry into the body portion 24. The needle port is connected to the body portion 24 with a thin wall of stainless steel tubing, and the needle ports are angled to provide a clear path to the bottom of the pyrotubes 34 a and 34 b. The tubing provides thermal resistance and allows the injector needles 46 a and 46 b to be maintained at a temperature lower than the temperature of the body portion 24. The injector needles 46 a and 46 b do not touch the sidewalls of the respective pyrotube 34 a and 34 b to maintain the injector needles 46 a and 46 b at a lower temperature than the detecting devices 26 a and 26 b. This provides for reliable methylating reagent metering and delivery. The body portion 24 is hot (approximately 230° C.), and the tubing keeps the needle port cool.

Each detecting device 26 a and 26 b includes a sample line 38 a and 38 b, respectively. During sample analysis, the sample lines 38 a and 38 b transport gaseous pyrolysis products from the pyrotubes 34 a and 34 b to a mass spectrometer 40 for analysis and identification. The sample lines 38 a and 38 b are stainless steel and lined with a fused quartz capillary liner. The quartz provides an inert medium for the reliable transport of the pyrolysis products to the mass spectrometer 40. The quartz is also resistant to the corrosive methylating reagent injected into the pyrotubes 34 a and 34 b during analysis. The quartz capillary liner may be positioned near the bottom of each pyrotube 34 a and 34 b to maximize the collection efficiency of the pyrolysis products.

Returning to FIG. 2, a spring loaded pyrotube lifting device 48 biases the pyrotubes 34 a and 34 b against the body portion 24 to prevent any clearance between a top edge 50 of the pyrotubes 34 a and 34 b and the body portion 24. Pushing the pyrotubes 34 a and 34 b fully home ensures that the gap between the pyrolyzer internal vertex and the ends of the pyrolyzer inlet tubes 30 a and 30 b remains constant. The top edge 50 is fire-polished to reduce internal stresses to prevent fractures when pushed against the body portion 24.

A cylindrical ferrule 52 provides a compression seal between the pyrotubes 34 a and 34 b and the body portion 24. The ferrule 52 is sold under the trademark VESPEL® and sold by E. I. Du Pont De Nemours and Company of Wilimington, Del. A second spring-loaded ferrule-lifting device 54 concentric with the pyrotube lifting device 48 applies pressure to the ferrule 52 to provide a pneumatic seal. The ferrule-lifting device 54 operates independently of the pyrotube lifting device 48. The ferrule-lifting device 54 applies a greater amount of force to the ferrule 52 than the pyrotube-lifting device 48 applies to the pyrotubes 34 a and 34 b. The pyrotube lifting device 48 pushes the pyrotubes 34 a and 34 b against the ferrule 52 to form a seal. A step 56 around the lower edge of the ferrule 52 engages a notch of the ferrule-lifting device 54. The ferrule-lifting device 54 can be lowered to remove the ferrule 52 from the biological agent detector 20.

A cylindrical heater 58 a and 58 b closely contoured to the bottom of the pyrotubes 34 a and 34 b provides heat during pyrolysis. A blower (not shown) provides forced air-cooling through ducts at the end of pyrolysis to cool the pyrotubes 34 a and 34 b in preparation for the next aerosol collection cycle.

The mass spectrometer 40 includes a preprogrammed list of molecular patterns. Each molecular pattern on the list is associated with a specific biological agent. If any biological agents are present the sample, the mass spectrometer 40 identifies the biological agents by comparing the identified molecular pattern to the list of molecular patterns preprogrammed into the mass spectrometer 40. After a match is found, the mass spectrometer 40 identifies the associated biological agent.

One detecting device 26 a and 26 b operates in a sample collection mode when the other detecting device 26 a and 26 b operates in a sample analysis mode. After a predetermined amount of time, the roles of the detecting devices 26 a and 26 b are reversed. Therefore, the biological agent detector 20 continuously collects an air sample for analysis. The mode-switching phase lasts approximately 0.5 seconds. During the change-over period, air is still collected. Preferably, the predetermined amount of time is one minute. However, one skilled in the art would know what predetermined amount of time to employ.

As shown in FIG. 4A, the detecting device 26 a operates in the sample collection mode, and the detecting device 26 b operates in the sample analysis mode. The concentrated aerosol outlet 62 of the bio-concentrator 22 aligns with the detecting device 26 a. An exhaust pump 78 a draws air through the exhaust line 36 a for exhausting to the atmosphere. When the exhaust pump 78 a is active, the exhaust pump 78 b of the detecting device 26 b is inactive and does not exhaust air through the exhaust line 36 b. The duration of the sample collection is programmable by software. In one example, the sample is collected for approximately 1 to 5 minutes. One skilled in the art would know how long to collect the sample. A control 87 controls the exhaust pumps 78 a and 78 b and the motor 70.

The sample lines 38 a and 38 b meet at a point, and an outlet line 120 extends from the point and directly to the mass spectrometer 40 to define a “T.” When detecting device 26 a is in sample collection mode and the detecting device 26 b is in sample analysis mode, the pressure in detecting device 26 a is lower than the ambient, and the pressure in the detecting device 26 b is greater than the ambient. There is a net flow of gas from the detecting device 26 b to the detecting device 26 a.

While the detecting device 26 a is operating in the sample collection mode, a sample previously collected in the pyrotube 34 b of the detecting device 26 b is analyzed. The methylating reagent is added to the sample in the pyrotube 34 b through the injector needle 46 b from the reagent source 44 b to split open the cells of any bacterial agents in the sample and neutralize and derivatize the organic molecules of the biological agents to form more volatile molecules. The sample in the pyrotube 34 b is then pyrolyzed or heated by the heater 58 b to form gas molecules. The sample can be pyrolyzed for approximately 1.5 minutes. However, one skilled in the art would know how long to pyrolyze the sample collected in the pyrotube 34 b.

The pyrolysis products are directed from the detecting device 26 b and along the sample line 38 b, the outlet line 120, and directly to the mass spectrometer 40. The detecting device 26 b has an internal pressure that is greater than the internal pressure of the detecting device 26 a, directing the pyrolysis products from the detecting device 26 b to the outlet line 120 and to the mass spectrometer 40. Approximately 1 milliliter per minute of the gas molecules are provided to the mass spectrometer 40.

After the predetermined amount of time, the detecting devices 26 a and 26 b switch functions. That is, the detecting device 26 a then operates in the sample analysis mode, and the detecting device 26 b operates in the sample collection mode. The motor 70 rotates the turntable 60 such that the concentrated exhaust outlet 62 of the bio-concentrator 22 aligns with the inlet tube 30 b of the detecting device 26 b to collect a sample of air in the pyrotube 34 b, as shown in FIG. 4B. The sample of air previously collected in the pyrotube 34 a of the detecting device 26 a is then analyzed.

When the detecting device 26 b operates is in sample collection mode and the detecting device 26 a operates in the sample analysis mode, the pressure in the detecting device 26 b is lower than the pressure in the detecting device 26 a. There is a net flow of gas from the detecting device 26 a to the detecting device 26 b. The gases from the detecting device 26 a flows through the “T” and along the outlet line 120 to be directly sampled by the mass spectrometer 40. That is, the pyrolysis products flow along the sample line 38 a and along the outlet line 120 to the mass spectrometer 40.

The biological agent detector 20 of the present invention can be used to detect biological agents used in biological warfare, such as anthrax. However, it is to be understood that other biological agents can be detected. Any substance having a unique pattern of fatty acids that can be identified by the mass spectrometer 40 can be detected and identified.

Although detecting devices 26 a and 26 b are disclosed as being two separate devices, it is to be understood that the detecting devices 26 a and 26 b could be integrated into a single assembly. In addition, other types of biological agent detectors could be used in place of the pyrolysis/mass spectrometer detector described herein.

The foregoing description is only exemplary of the principles of the invention. Many modifications and variations of the present invention are possible in light of the above teachings. The preferred embodiments of this invention have been disclosed, however, so that one of ordinary skill in the art would recognize that certain modifications would come within the scope of this invention. It is, therefore, to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described. For that reason the following claims should be studied to determine the true scope and content of this invention. 

1. A method of detecting a biological agent in an air sample, the method comprising the steps of: flowing a first aerosol sample into a first detecting device during a sample collection mode, wherein an air sample is collected during the sample collection mode; identifying biological agents in a second aerosol sample collected in a second detecting device during a sample analysis mode, wherein the air sample is analyzed and biological agents in the air sample are identified, and the step of identifying the biological agents in the second aerosol sample occurs during the step of flowing the first aerosol sample into the first detecting device; flowing a third aerosol sample into the second detecting device after the step of identifying the biological agents in the second aerosol sample; and identifying the biological agents in the first aerosol sample collected in the first detecting device, wherein the step of identifying the biological agents in the first air sample in the first detecting device occurs during the step of then flowing the third aerosol sample into the second detecting device.
 2. The method as recited in claim 1 wherein the step of identifying the biological agents in the second aerosol sample includes heating the second aerosol sample during the sample analysis mode.
 3. The method as recited in claim 1 wherein the step of identifying the biological agents in the second aerosol sample includes transporting the second aerosol sample to a chemical analyzing device for analysis during the sample analysis mode.
 4. The method as recited in claim 1 including the step of injecting a methylating reagent into the second air sample during the sample analysis mode.
 5. The method as recited in claim 4 wherein the methylating reagent is TMAH.
 6. The method as recited in claim 1 including the step of exhausting a portion of the first air sample drawn into the first detecting device during the sample collection mode.
 7. The method as recited in claim 1 including the step of concentrating the first air sample into the first aerosol sample prior to the step of flowing the first aerosol sample into the first detecting device.
 8. The method as recited in claim 7 wherein the step of concentrating the first air sample to the first aerosol sample is performed by a bio-concentrator shared by the first detecting device and the second detecting device.
 9. The method as recited in claim 8 including the step of moving the bio-concentrator between a first position where the first aerosol sample is directed to the first detecting device and a second position where the second aerosol sample is directed to the second detecting device. 